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Although disturbances in body function of animals can be measured to determine whether a state of stress may exist, there is growing interest in finding ways to assess their emotional status as an indicator of good or bad welfare status. Generally it is easier to determine poor states of well-being than positive ones. For grazing ruminants some indicators of well-being include absence of illness, good growth and productivity, and longevity. Motion detectors can provide automated remote monitoring of behavior and it is likely that there will be advances in the interpretation software to increase the utility of this technology for assessing well-being. Cortisol levels in body fluids, feces and pelage are prominent as a marker of poor animal welfare, but like many of the other objective measures that are used, are not wholly reliable at the individual animal level. These other measures include: plasma serotonin, heart rate variation, infra-red thermography, cytokines, salivary alpha amylase, and acute phase proteins. Use of automated facial expression recognition may supplement electrophysiological recording as means to quantify the pain experience of animals. Although the measures described in the literature do not necessarily provide the final answer for determination of welfare in grazing ruminants, they all have some merit and deserve further investigation.

Coumestrol produced by alfalfa (Medicago sativa L.) can reduce the ovulation rate of ewes. This study isolated agronomic factors that affect coumestrol levels in alfalfa. Fungal diseases explained most differences in coumestrol. Alfalfa inoculated with Stemphylium vesicarium contained 169 ± 25.1 mg kg⁻¹ dry matter (DM) compared with 3.4 ± 0.84 mg kg⁻¹ DM in controls. However, in field-grown alfalfa there was a low relationship (R² = 0.388) between coumestrol content and visual fungal damage score which indicates a need for alternative methods of prediction. In the field, a range of cultivars all reached coumestrol levels reported to pose a risk to ewe reproductive performance (>25 mg kg⁻¹ DM). In S. vesicarium-inoculated leaves, the modern cultivar ‘Stamina 5’ had 396 ± 82.4 mg kg⁻¹ DM compared with the 40-yr-old industry standard ‘Wairau’ at 143 ± 35.6 mg kg⁻¹ DM. Flowering in isolation did not produce a coumestrol response, with increases detected or not detected simultaneously in vegetative and flowering plants. Pea aphids had a minor effect on coumestrol content. When alfalfa was subjected to ~5 aphids per stem for 4 wk coumestrol increased from 2.4 ± 0.39 to 5.3 ± 0.65 mg kg⁻¹ DM. This study highlights fungal pathogens traditionally not considered problematic in alfalfa stands due to limited effects on stand persistence and yield, as important causal agents of elevated coumestrol levels. Plant breeding to reduce their impact is recommended.

Mutations in the CLN5 gene cause the fatal, pediatric, neurodegenerative disease CLN5 neuronal ceroid lipofuscinosis. Affected children suffer progressive neuronal loss, visual failure and premature death. Presently there is no treatment. This study evaluated dual intracerebroventricular (ICV) and intravitreal (IVT) administration of a self-complementary adeno-associated viral vector encoding ovine CLN5 (scAAV9/oCLN5) into CLN5 affected sheep (CLN5−/−) at various disease stages. CLN5 disease progression was slowed in pre-symptomatic sheep who received a moderate dose of scAAV9/oCLN5, whilst a higher ICV dose treatment in early and advanced symptomatic animals delayed or halted disease progression. Intracranial (brain) volume loss was attenuated in all treatment cohorts, and visual function was also sustained in both the early and advanced symptomatic treated sheep over the 24-month duration of the study. Robust CLN5 protein expression was detected throughout the brain and spinal cord, and improvements in central nervous system and retinal disease correlates were observed. These findings hold translational promise for extending and improving the quality of life in both pre-symptomatic and symptomatic CLN5 patients, and prompted the initiation of the first in-human Phase I/II clinical trial testing ICV/IVT administration of scAAV9 encoding human CLN5 (https://clinicaltrials.gov/; NCT05228145).

Introduction: The neuronal ceroid lipofuscinoses (NCLs; Batten disease) are a group of fatal neurodegenerative lysosomal storage diseases of children caused by various mutations in a range of genes. Forms associated with mutations in two of these, CLN5 and CLN6, are being investigated in well-established sheep models. Brain atrophy leading to psychomotor degeneration is among the defining features, as is regional progressive ossification of the inner cranium. Ongoing viral-mediated gene therapy trials in these sheep are yielding encouraging results. In vivo assessment of brain atrophy is integral to the longitudinal monitoring of individual animals and provides robust data for translation to treatments for humans. Methods: Computed tomography (CT)-based three-dimensional reconstruction of the intracranial volume (ICV) over time reflects the progression of cortical brain atrophy, verifying the use of ICV measurements as a surrogate measure for brain size in ovine NCL. Results: ICVs of NCL-affected sheep increase for the first few months, but then decline progressively between 5 and 13 months in CLN5⁻/⁻ sheep and 11–15 months in CLN6⁻/⁻ sheep. Cerebral ventricular volumes are also increased in affected animals. To facilitate ICV measures, the radiodensities of ovine brain tissue and cerebrospinal fluid were identified. Ovine brain tissue exhibited a Hounsfield unit (HU) range of (24; 56) and cerebrospinal fluid a HU range of (−12; 23). Conclusions: Computed tomography scanning and reconstruction verify that brain atrophy ovine CLN5 NCL originates in the occipital lobes with subsequent propagation throughout the whole cortex and these regional differences are reflected in the ICV loss.

Inefficient nitrogen (N) use from pastoral dairy production systems has resulted in environmental degradation, as a result of excessive concentrations of urinary N excretion leaching into waterways and N₂O emissions from urination events into the atmosphere. The objectives of this study were to measure and evaluate the total N balance of lactating dairy cows selected for milk urea N concentration breeding values (MUNBVs) consuming either a 100% perennial ryegrass (Lolium perenne L.) or 100% plantain (Plantago lanceolata L.) diet. Sixteen multiparous lactating HolsteinFriesian × Jersey cows divergent for MUNBV were housed in metabolism crates for 72 h, where intake and excretions were collected and measured. No effect of MUNBV was detected for total N excretion; however, different excretion characteristics were detected, per urination event. Low MUNBV cows had a 28% reduction in the concentration of urinary urea nitrogen (g/event) compared to high MUNBV cows when consuming a ryegrass diet. Cows consuming plantain regardless of their MUNBV value had a 62% and 48% reduction in urinary urea nitrogen (g/event) compared to high and low MUNBV cows consuming ryegrass, respectively. Cows consuming plantain also partitioned more N into faeces. These results suggest that breeding for low MUNBV cows on ryegrass diets and the use of a plantain diet will reduce urinary urea nitrogen loading rates and therefore estimated nitrate leaching values, thus reducing the environmental impact of pastoral dairy production systems.

We report a novel method of investigating foraging in marine mammals based on detecting biomarkers of strictly dietary origin in blood. Arsenobetaine (AsB), the arsenic analogue of the osmolyte glycine betaine, and trimethylamine N-oxide (TMAO), an osmolyte used by marine fish and invertebrates, were measured in plasma of lactating Weddell seals Leptonychotes weddellii during the postpartum fast and at different stages of lactation. Plasma dietary biomarker concentrations were low in early lactation (<14 d postpartum [d p.p.]; AsB: 5.5 ± 2 ppb As, TMAO: 66 ± 20 μmol l-1) and increased 10-fold in late lactation (≥27 d p.p.; AsB: 57 ± 17 ppb As, TMAO: 685 ± 199 μmol l-1). In lactating females (n = 6) monitored longitudinally, plasma TMAO remained low for the first 3 wk p.p. Increases in AsB concentrations observed in late lactation were closely correlated (Pearson correlation r = 0.96, p < 0.01, n = 6) with increases in plasma insulin-like growth factor 1 (IGF1), a growth factor known to reflect nutritional status in other mammals. Two seals outfitted with time-depth recorders began regular deep diving at different times p.p. and showed corresponding differences in biomarker concentrations. Our results indicate that (1) most lactating Weddell seals at the study site (70 % of 20 seals studied) feed after 3 to 4 wk p.p., (2) individual lactating Weddell seals may forage as early as 9 d p.p., and (3) some Weddell seals may consume little or no food throughout lactation. Dietary biomarkers provide a simple method for distinguishing feeding and fasting in free-living marine mammals. © Inter-Research 2005.

To identify separate effects of gender, castration and exogenous oestrogen on growth, castrated lambs of both sexes and entire male lambs (n = 8) were implanted subcutaneously with three sizes of oestradiol-17β implants, or not implanted, and grazed on ryegrass and white clover pasture for 180 days. A group of non-implanted entire female lambs (n = 8) was run together with the others. Non-implanted entire male lambs grew faster, had heavier heads, less internal, non-carcass fat and more protein and less fat and water in the carcass than non-implanted entire females. In addition, they had higher 12th vertebral spine, thicker tibia, and heavier and larger humerus than entire female lambs. Castration of male lambs reduced live-weight gain, weight of head and content of protein in the carcass whereas it increased carcass fat content. In addition, it caused lengthening of cannon bones and reduced height of 12th vertebral spine and length of tibia. In females, gonadectomy increased height of 12th vertebral spine and diameter to length ratio of the radius. Oestradiol treatment increased live-weight gain, reduced total internal and carcass fat, and increased water and protein content of the carcass in gonadectomized animals of either sex, and increased weight of carcass and head in spayed ewe lambs. Oestradiol treatment inhibited longitudinal growth of cannon bones and stimulated that of vertebral column and ribs, but had little effect on the dimensions of limb bones apart from increasing their diameter. Oestradiol treatment had no effect on muscle length but increased muscle girth and weight, except for m. splenius in ram lambs where muscle weight was reduced. Effects of oestradiol on skeletal measurements in most cases were linearly related to dose of oestradiol. It was concluded that the variable effects of sex steroids on the skeleton were related to the differential pattern of skeletal maturation. In early maturing bones acceleration of the growth process by an exogenous sex steroid caused elongation to cease prematurely, whereas in late-maturing bones the acceleration effect on elongation did not result in premature cessation. This observation may explain the often contradictory reports in the literature on the effects of sex steroids on linear growth of bone.